Chlamydomonas reinhardtii NADP-linked glyceraldehyde-3-phosphate dehydrogenase contains the cysteine residues identified as potentially domain-locking in the higher plant enzyme and is light activated

The chloroplastic glyceraldehyde-3-P dehydrogenase (EC 1.2.1.13) of the green alga Chlamydomonas reinhardtii is reductively light activated. Homology modeling indicates that the only potential disulfide-forming cysteine residues in this enzyme are the same cysteine residues suggested to be responsible for redox-sensitivity of the higher plant enzyme (Li D, Stevens FJ, Schiffer M and Anderson LE (1994) Biophys J 67: 29–35). Apparently, the three additional cysteines in the higher plant enzyme are not necessary for light activation. The putative regulatory cysteines are juxtaposed across the domain interface and when oxidized will crosslink the domains. This would be expected to interfere with interdomain movement and catalysis. This is the first report of reductive light activation of this enzyme in a green alga.     

The oligotrophic peatlands of Western Siberia-the largest peino-helobiome in the world

Summary The paper summarizes the vegetation pattern of the largest swamp (peat bog) area in the world. The core area covers about 800×1800 km (NS x EW) in West Siberia between the Ural mountains and the river Yenisey. The core area is one gigantic oligotrophic swamp (Peino-Helobiome acc), the new classification suggested in Walter 1976, see also summary in Vegetatio 32, 1976, pp. 75–81. A map of the entire area is presented.Intensive recent field studies were executed by the geobotanical-soil science faculty of the University of Moscow and the first five publication under the editorship of A. I. Popov (1971–1975) are the basis for this paper. The original papers are in Russian, still unavailable for most western scientists.The bog genetics was stratigraphically analyzed. The analysis revealed that the majority of the Northern bogs started in taiga forests. The peat in these areas is now 4–7 m deep. About 60% of the peat consists of the mixed (Sphagnum) fuscum type deposit. Peat type profiles are shown in Figure 4. The figure illustrates the different peat deposits in relation to depth as well as to surface pattern of strings and hollows.The surface vegetation shows the pattern typical for many northern peat bogs on very slightly inclined land: parallel strings of peat moss and Pinus trees alternating with hollows filled with rain water and hydrophytic Sphagna (Sph. dusenii, Sph. balticum). A cross section through this vegetation pattern is given in Figure 5. Details of the pattern: strings and hollows are given in Chapter 4 of the paper.The second chapter of this paper deals with regression phenomena in the vegetation development due to excessive water logging caused by the development of the strings. Under such conditions black water lakes are formed which grow in size through the erosive action of wind and waves on the expense of the peat moss surface.Bog development in the middle and northern Taiga zone of the biome are discussed in Section 3. The hydrology of this area is ruled by the seasonal fluctuation of water level. The amplitude of the Ob river amounts to 12 m with the amplitude of the tributaries ranging around 4–6 m. The annual flood is aggravated in spring by huge ice packages. The entire area is for extended periods one great lake which dissects into hundreds of thousands of little lakes during the low water season.In some parts of this area local drainage is responsible for the development of small forest patches. The crown cover in this forest is reported up to 50%; the species composition changes from Pinus sibirica, P. sylvestris, Picea spec. near the drainage creeks to Pinus sylvestris f. willkommii and Betula in the drained upland portions.In this area the general peat bog is eutrophic because of the high influence of the flood. The local development of oligotrophic raised bogs is frequent, however. These bogs are obviously a successional stage in series that leads eventually to a boreal forest.Section 5 deals with the vegetation typology of the entire area. Three different subdivisions of the Helobiome are distinguished, with several regional subdivisions in each. The principle for the biome division is the environmental change from oligotrophic water on the northern lower end of the watershed to the eutrophic water condition further south in the forest steppe zone. In the area of the southern eutrophic biome are forest islands Ryami and even the first salt soil patches noticeable especially in the northern steppe zone. This indicates the regional transition to the Halo-Helobiome. Some vegetational and floristic details for the subdivisions are summarized in this chapter.Supplement to H. Walter: The vegetation of Eastern Europe, North and Central Asia, 1974.Translated by: Stephen S. Talbot, Forest Management Institute, 396 Cooper Street, Ottawa, Ontario, Canada K1A OH3, and Runhild E. Eessell, 54 Spindle Road Hicksville, New York 11801, USA.

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Where did Marenzelleria spp. (Polychaeta: Spionidae) in Europe come from?

Abundant populations of Marenzelleria spp. were reported for the first time in the North Sea during the late 1970s and then in the Baltic Sea in 1985. Genetic analysis by means of allozyme electrophoresis and sequencing of a segment of mitochondrial 16srDNA showed that two different genetic types or sibling species of Marenzelleria were present in Europe. Marenzelleria Type I is found only in the North Sea, whereas Type II has been found in both the North Sea and the Baltic Sea. The North Sea animals (Type I) correspond to Type I specimens found in North American coastal waters between Nova Scotia and Cape Henlopen (Delaware), while Marenzelleria Type II from the Baltic Sea correspond to Marenzelleria type II animals from the Arctic (Tuktoyaktuk Harbor, Northwest Territories, Canada), New Hampshire and coastal waters between Chesapeake Bay southward to the Ogeechee River (Georgia). Human mediated introduction (by shipping) and natural range expansion are discussed as the possible causes of these virtually simultaneous invasions by two sibling species. Marenzelleria Type I appears to colonize habitats with a higher salinity and/or in which salinities tend to fluctuate considerably. The osmolality of the coelomic fluid after acclimation to various salinities between 0.25 and 18 is the same for both sibling species/genetic types. Although the two types do not differ in respect of hyperosmoregulation (<10), differences may exist in their cell volume regulation or its time course in the almost isoosmotic range at salinities >10.     

Subtractive hybridization of cDNA from small amounts of plant tissue

A subtractive hybridization method is described that allows the generation of a subtractive gene library from small amounts of plant or other eukaryotic tissues. The method uses paramagnetic oligo-dT beads to capture poly-adenylated mRNA and to synthesize the complementary cDNA on a solid support. The use of magnetic beads facilitates the change of reaction buffers and the removal of primers and minimizes yield losses. Subtracted material obtained from this method can either be cloned directly or used to screen a specific library.     

Reformable intramolecular cross-linking of the N-terminal domain of heparin cofactor II: effects on enzyme inhibition.

The crystal structure of a heparin cofactor II (HCII)-thrombin Michaelis complex has revealed extensive contacts encompassing the N-terminal domain of HCII and exosite I of the proteinase. In contrast, the location of the N-terminal extension in the uncomplexed inhibitor was unclear. Using a disulfide cross-linking strategy, we demonstrate that at least three different sites (positions 52, 54 and 68) within the N terminus may be tethered in a reformable manner to position 195 in the loop region between helix D and strand s2A of the HCII molecule, suggesting that the N-terminal domain may interact with the inhibitor scaffold in a permissive manner. Cross-linking of the N terminus to the HCII body does not strongly affect the inhibition of alpha-chymotrypsin, indicating that the reactive site loop sequences of the engineered inhibitor variants, required for interaction with one of the HCII target enzymes, are normally accessible. In contrast, intramolecular tethering of the N-terminal extension results in a drastic decrease of alpha-thrombin inhibitory activity, both in the presence and in the absence of glycosaminoglycans. Treatment with dithiothreitol and iodoacetamide restores activity towards alpha-thrombin, suggesting that release of the N terminus of HCII is an important component of the multistep interaction between the inhibitor and alpha-thrombin.     

pEffects of UV radiation on the ultrastructure of several red algae

The effect of ultraviolet (UV) radiation on the ultrastructure of four red algae, the endemic Antarctic Palmaria decipiens (Reinsch) Ricker and Phycodrys austrogeorgica Skottsberg, the Arctic‐cold temperate Palmaria palmata (Linnaeus) O. Kuntze and the cosmopolitan Bangia atropurpurea (Roth) C. Agardh was studied. All four species showed a formation of ‘inside‐out’ vesicles from the chloroplast thylakoids upon exposure to artificial UV‐radiation. In P. decipiens, most vesicles were developed after 8 h and in P. palmata, after 48 h of UV exposure. In B. atropurpurea, vesi‐culation of thylakoids was observed after 72 h of UV irradiation. In Ph. austrogeorgica, the chloroplast envelope and thylakoid membranes were damaged and the phycobilisomes became detached from the thylakoids after 12 h of UV exposure. Ultraviolet‐induced changes in the membrane structure of mitochondria were observed in P. decipiens and P. palmata. However, in P. decipiens they were reversible as was the damage in chloroplast fine structure after 12 h of UV treatment. Protein crystals in Ph. austrogeorgica showed degradation after exposure to UV radiation. Different methods of fixation and embedding macroalgal material are discussed. These findings give insight into the fine structural changes which occur during and after UV exposure and indicate a relationship between the species dependent sensitivity to UV‐exposure and the depth distribution of the different species.     

Sensitive In Situ Monitoring of a Recombinant Bioluminescent Yersinia enterocolitica Reporter Mutant in Real Time on Camembert Cheese

Bioluminescent mutants of Yersinia enterocolitica were generated by transposon mutagenesis using a promoterless, complete lux operon (luxCDABE) derived from Photorhabdus luminescens, and their production of light in the cheese environment was monitored. Mutant B94, which had the lux cassette inserted into an open reading frame of unknown function was used for direct monitoring of Y. enterocolitica cells on cheeses stored at 10°C by quantifying bioluminescence using a photon-counting, intensified charge-coupled device camera. The detection limit on cheese was 200 CFU/cm². Bioluminescence of the reporter mutant was significantly regulated by its environment (NaCl, temperature, and cheese), as well as by growth phase, via the promoter the lux operon had acquired upon transposition. At low temperatures, mutant B94 did not exhibit the often-reported decrease of photon emission in older cells. It was not necessary to include either antibiotics or aldehyde in the food matrix in order to gain quantitative, reproducible bioluminescence data. As far as we know, this is the first time a pathogen has been monitored in situ, in real time, in a “real-product” status, and at a low temperature.     

Cloning and characterization of the mammalian-specific nicolin 1 gene (NICN1) encoding a nuclear 24 kDa protein.

We have identified a novel mammalian gene, termed nicolin 1 gene (NICN1), that is present in human, dog and mouse, whereas it is absent from the available genome sequences of nonmammalian organisms. The NICN1 gene consists of six exons and spans about 6 kb of genomic DNA. It encodes a 213 amino acid protein that does not belong to any known protein family. Experiments using green fluorescent protein (GFP)-tagged nicolin 1 fusion proteins indicate that nicolin 1 is a nuclear protein. Northern analysis and semiquantitative RT-PCR demonstrated that the 2.5 kb NICN1 mRNA is expressed in a tissue-specific manner. The highest NICN1 expression levels are found in brain, testis, liver, and kidney. On the other hand the NICN1 expression is weak in spleen, leukocytes, small intestine and colon. The NICN1 gene is also expressed during development.